منابع مشابه
Generation of families of construct variants using golden gate shuffling.
Current standard cloning methods based on the use of restriction enzymes and ligase are very versatile, but are not well suited for high-throughput cloning projects or for assembly of many DNA fragments from several parental plasmids in a single step. We have previously reported the development of an efficient cloning method based on the use of type IIs restriction enzymes and restriction-ligat...
متن کاملAssembly of Designer TAL Effectors by Golden Gate Cloning
Generation of customized DNA binding domains targeting unique sequences in complex genomes is crucial for many biotechnological applications. The recently described DNA binding domain of the transcription activator-like effectors (TALEs) from Xanthomonas consists of a series of repeats arranged in tandem, each repeat binding a nucleotide of the target sequence. We present here a strategy for en...
متن کاملMobius Assembly: A Versatile Framework For Golden Gate Assembly
Golden Gate Assembly is a powerful synthetic biology tool, which utilizes Type IIS enzymes for unidirectional assembly of multiple DNA fragments. The simplicity of its DNA assembly and the exchangeability of standard parts greatly facilitate the generation of combinatorial assembly libraries. Currently there are two popular Golden Gate Assembly frameworks that allow multigene augmentation (MoCl...
متن کاملGolden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes
We have developed a protocol to assemble in one step and one tube at least nine separate DNA fragments together into an acceptor vector, with 90% of recombinant clones obtained containing the desired construct. This protocol is based on the use of type IIs restriction enzymes and is performed by simply subjecting a mix of 10 undigested input plasmids (nine insert plasmids and the acceptor vecto...
متن کاملWhere are they now? A follow-up study of suicide attempters from the Golden Gate Bridge.
The Golden Gate Bridge is currently the number one suicide location in the world. From the opening day, May 18, 1937 to April 1, 1978, there have been 625 officially reported suicide deaths and perhaps more than 200 others which have gone unseen and unreported. Proposals for the construction of a hardware antisuicide barrier have been challenged with the untested contention that "they'll just g...
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ژورنال
عنوان ژورنال: Koedoe
سال: 1968
ISSN: 2071-0771,0075-6458
DOI: 10.4102/koedoe.v11i1.755